Cryostat for cutting flash-frozen tissue sections

所要求的设备是一台徕卡低温恒温器，用于切割先前快速冷冻组织的薄片。Gardiner的nserc资助的研究的一个主要部分是确定蛋白质的水平，以及为这些蛋白质编码的信使rna，在老鼠的神经元中控制后肢肌肉，即α运动神经元。另一个研究方向涉及测量肥胖和糖尿病对大鼠血流的影响及其在各种不同组织中的调节。在这两种情况下，从大鼠身上取出的组织都必须迅速冷冻，然后在大约零下20摄氏度的温度下切片，以便随后对组织进行处理，使我们感兴趣的元素可视化。Stecina的nserc资助的研究项目包括鉴定成年啮齿动物神经系统中的脊髓中间神经元及其功能网络。在几乎所有这些实验中，除了电生理学研究之外，还需要事后验证大脑中刺激电极的位置。此外，对于神经元间网络的电生理表征，神经解剖学方法，如活动依赖标记，脊髓中记录位点的识别也在每次实验后实际使用。此外，每次电生理实验后采集的脊髓和脑组织切片的切割需要经常使用低温恒温器（每周至少1 - 2天）。对于Cowley来说，nserc资助的关于行走和平衡的神经基础的研究需要使用低温恒温器进行病变验证，研究涉及到中间神经元和运动神经元的逆行标记，以及与神经递质受体分布相关的免疫组织化学。我们目前使用的低温恒温器已经使用了15年，使用频繁，而且正在损坏。购买所要求的设备将使所有这些研究项目能够不间断地继续下去，这些研究项目对我们理解我们如何运动、参与运动的神经网络如何受到高级中枢的控制、环境如何影响神经和其他组织的基因表达和功能都有影响。当目前使用的低温恒温器停止正常工作时，我们预计这将随时发生，因为它的年龄，无法获得更换部件，以及目前的大量使用，三位研究者的研究计划将受到严重损害，以至于他们各自的研究计划的整个部分将无法完成。

The equipment requested is a Leica cryostat for cutting of thin sections of previously rapidly-frozen tissue. A major part of Gardiner's NSERC-funded research involves determining the level of proteins, and of the messenger RNAs which code for these proteins, in the neurons in rats which control the hindlimb muscles, the alpha-motoneurons. Another research direction involves measuring the effects of obesity and diabetes in rats on blood flow and its regulation in a variety of different tissues. In both cases, tissues removed from the rat must be rapidly frozen, and later sectioned at a temperature of about minus 20 degrees C, to treat the tissues subsequently to visualize the elements we are interested in. Stecina's NSERC-funded research program involves the identification of spinal interneurons and their functional networks in the adult rodent nervous system. In almost all of those experiments, in addition to the electrophysiological studies, verification of the placement of stimulation electrodes in the brain is required post-hoc. Also, for the electrophysiological characterization of interneuronal networks, neuroanatomical methods, such as activity dependent labelling, identification of recording sites in the spinal cord are also used practically after each of the performed experiment. In addition, the cutting of thin sections from spinal cord and brain tissue harvested after each electrophysiological experiment warrants the use of a cryostat on a frequent basis (minimum 1 - 2 days per week). For Cowley, NSERC-funded investigations of the neural basis of stepping and balance require the use of a cryostat for lesion verification, studies involving retrograde labeling of interneurons and motoneurons, as well as for immunohistochemistry related to the neurotransmitter receptor distribution. We currently use a cryostat that is 15 years old, which is heavily used, and is breaking down. The purchase of the requested equipment will allow all of these research programs, which have implications for our understanding of how we move, how the neural networks involved in movements are controlled by higher centres and how the environment influences the gene expression and function of nervous and other tissues, to continue without interruption. When the currently-used cryostat ceases to function properly, which we expect will occur at any time due to its age, the unavailability of replacement parts, and current heavy usage, the research programs of the three investigators will be severely compromised, such that entire portions of their respective research programs will be impossible to complete.